Root-knot nematode chemotaxis is positively regulated by l-galactose sidechains of mucilage carbohydrate rhamnogalacturonan-I

Root-knot nematodes (RKNs) are plant parasites and major agricultural pests. RKNs are thought to locate hosts through chemotaxis by sensing host-secreted chemoattractants; however, the structures and properties of these attractants are not well understood. Here, we describe a previously unknown RKN attractant from flaxseed mucilage that enhances infection of Arabidopsis and tomato, which resembles the pectic polysaccharide rhamnogalacturonan-I (RG-I). Fucose and galactose sidechains of the purified attractant were found to be required for attractant activity. Furthermore, the disaccharide α-l-galactosyl-1,3-l-rhamnose, which forms the linkage between the RG-I backbone and galactose sidechains of the purified attractant, was sufficient to attract RKN. These results show that the α-l-galactosyl-1,3-l-rhamnose linkage in the purified attractant from flaxseed mucilage is essential for RKN attraction. The present work also suggests that nematodes can detect environmental chemicals with high specificity, such as the presence of chiral centers and hydroxyl groups.

Attraction and penetration of Musa by the burrowing nematode, Radopholus similis, in an autotrophic bipartite culture system

Summary Our objective was to discover the stages (pre- or post-infection) in which the resistance to burrowing nematode (Radopholus similis) occurs in two resistant banana (Musa spp.) cultivars. An autotrophic in vitro culture system was used to compare R. similis migration towards, and penetration into, the banana roots. A new two-compartment autotrophic in vitro model system was developed using agar-based medium to examine the migration of R. similis to either the susceptible ‘Grande Naine’ or the resistant ‘Yangambi km5’ (‘Ykm5’), when both the Musa genotypes were present at equal distance. The autotrophic in vitro model system was advantageous, because it supported continuous root growth due to the actively photosynthesising shoots growing in the open air, while the in vitro root conditions make it possible to observe and assess the nematode chemotaxis in the transparent medium. Significantly fewer nematodes migrated towards the resistant ‘Ykm5’ plants when compared to both the susceptible ‘Grande Naine’, and another resistant cultivar, ‘Saba’, at 1 h after infection. This signals a possibility of a lower concentration or different composition of nematode attractants in ‘Ykm5’ root exudates. No significant differences were observed in the percentage of R. similis that migrated towards the roots of the susceptible and resistant banana plants at 3, 4 and 6 h after inoculation. No significant differences were observed in the percentages of female penetration in the resistant and susceptible plant roots at 1 and 2 days after inoculation. The results of the two-compartment system confirmed that when a choice is given to migrate towards the resistant and susceptible genotypes, no differences were observed in the percentage of female migration towards both the genotypes.

Exploring Predatory Nematode Chemotaxis Using Low-Cost and Easy-to-Use Microfluidics

Symbiosis is a fascinating and diverse phenomenon. The study of symbiosis is important to understanding ecology, as it helps us understand relationships between organisms and provides insight into co-evolution, mutualism, adaptation, and survival. Ecological studies are challenging to implement in K-12 classrooms because they often require multiple organisms (often very different in size) and complex environments that are difficult to replicate accurately (e.g., soil composition, temperature, pH, and humidity). These factors can make it difficult to study quantitative changes in ecosystems. We developed an inexpensive, quantitative experiment for classrooms that can be used to explore important aspects of microbial symbiosis, pathogenesis, and ecology, and that helps support more investigations in this area of education. The experiment is low-cost, designed for K-12 teachers and students, uses common materials, and teaches students about the exciting relationships among bacteria, worms, and insects.

Chip Technologies for Screening Chemical and Biological Agents Against Plant-Parasitic Nematodes

Plant-parasitic nematodes cause substantial damage to agricultural crops worldwide. Long-term management of these pests requires novel strategies to reduce infection of host plants. Disruption of nematode chemotaxis to root systems has been proposed as a potential management approach, and novel assays are needed to test the chemotactic behavior of nematodes against a wide range of synthetic chemicals and root exudates. Two microfluidic chips were developed that measure the attraction or repulsion of nematodes to chemicals (“chemical chip”) and young plant roots (“root chip”). The chip designs allowed for chemical concentration gradients to be maintained up to 24 h, the nematodes to remain physically separate from the chemical reservoirs, and for images of nematode populations to be captured using either a microscope or a flatbed scanner. In the experiments using the chemical chips, seven ionic solutions were tested on second-stage juveniles (J2s) of Meloidogyne incognita and Heterodera glycines. Results were consistent with previous reports of repellency of M. incognita to a majority of the ionic solutions, including NH4NO3, KNO3, KCl, MgCl2, and CaCl2. H. glycines was found to be attracted to both NH4NO3 and KNO3, which has not been reported previously. A software program was written to aid in monitoring the location of nematodes at regular time intervals using the root chip. In experiments with the root chip, H. glycines J2s were attracted to roots of 3-day-old, susceptible (cultivar Williams 82) soybean seedlings, and attraction of H. glycines to susceptible soybean was similar across the length of the root. Attraction to resistant (cultivar Jack) soybean seedlings relative to the water only control was inconsistent across runs, and H. glycines J2s were not preferentially attracted to the roots of resistant or susceptible cultivars when both were placed on opposite sides of the same root chip. The chips developed allow for direct tests of plant-parasitic nematode chemotaxis to chemicals and roots with minimal human intervention.